Spinach hybrid SV8545VC and parents thereof

ABSTRACT

The invention provides seed and plants of spinach hybrid SV8545VC and the parent lines thereof. The invention thus relates to the plants, seeds and tissue cultures of spinach hybrid SV8545VC and the parent lines thereof, and to methods for producing a spinach plant produced by crossing such plants with themselves or with another spinach plant, such as a plant of another genotype. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of such plants, including the leaf and gametes of such plants.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the priority of U.S. Provisional Appl. Ser. No.62/024,380, filed Jul. 14, 2014, the entire disclosure of which isincorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to the field of plant breeding and, morespecifically, to the development of spinach hybrid SV8545VC, and parentspinach lines SMB66-1144F and SMBS012-1188M.

BACKGROUND OF THE INVENTION

The goal of vegetable breeding is to combine various desirable traits ina single variety/hybrid. Such desirable traits may include any traitdeemed beneficial by a grower and/or consumer, including greater yield,resistance to insects or pests, tolerance to environmental stress,better agronomic quality, higher nutritional value, growth rate andfruit properties.

Breeding techniques take advantage of a plant's method of pollination.There are two general methods of pollination: a plant self-pollinates ifpollen from one flower is transferred to the same or another flower ofthe same plant or plant variety. A plant cross-pollinates if pollencomes to it from a flower of a different plant variety.

Plants that have been self-pollinated and selected for type over manygenerations become homozygous at almost all gene loci and produce auniform population of true breeding progeny, a homozygous plant. A crossbetween two such homozygous plants of different genotypes produces auniform population of hybrid plants that are heterozygous for many geneloci. Conversely, a cross of two plants each heterozygous at a number ofloci produces a population of hybrid plants that differ genetically andare not uniform. The resulting non-uniformity makes performanceunpredictable.

The development of uniform varieties requires the development ofhomozygous inbred plants, the crossing of these inbred plants, and theevaluation of the crosses. Pedigree breeding and recurrent selection areexamples of breeding methods that have been used to develop inbredplants from breeding populations. Those breeding methods combine thegenetic backgrounds from two or more plants or various other broad-basedsources into breeding pools from which new lines and hybrids derivedtherefrom are developed by selfing and selection of desired phenotypes.The new lines and hybrids are evaluated to determine which of those havecommercial potential.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a spinach plant of thehybrid designated SV8545VC, the spinach line SMB66-1144F, orSMBS012-1188M. Also provided are spinach plants having all thephysiological and morphological characteristics of such a plant. Partsof these spinach plants are also provided, for example, includingpollen, an ovule, scion, a rootstock, a fruit, and a cell of the plant.

In another aspect of the invention, a plant of spinach hybrid SV8545VCand/or spinach lines SMB66-1144F and SMBS012-1188M comprising an addedheritable trait is provided. The heritable trait may comprise a geneticlocus that is, for example, a dominant or recessive allele. In oneembodiment of the invention, a plant of spinach hybrid SV8545VC and/orspinach lines SMB66-1144F and SMBS012-1188M is defined as comprising asingle locus conversion. In specific embodiments of the invention, anadded genetic locus confers one or more traits such as, for example,herbicide tolerance, insect resistance, disease resistance, and modifiedcarbohydrate metabolism. In further embodiments, the trait may beconferred by a naturally occurring gene introduced into the genome of aline by backcrossing, a natural or induced mutation, or a transgeneintroduced through genetic transformation techniques into the plant or aprogenitor of any previous generation thereof. When introduced throughtransformation, a genetic locus may comprise one or more genesintegrated at a single chromosomal location.

The invention also concerns the seed of spinach hybrid SV8545VC and/orspinach lines SMB66-1144F and SMBS012-1188M. The spinach seed of theinvention may be provided as an essentially homogeneous population ofspinach seed of spinach hybrid SV8545VC and/or spinach lines SMB66-1144Fand SMBS012-1188M. Essentially homogeneous populations of seed aregenerally free from substantial numbers of other seed. Therefore, seedof hybrid SV8545VC and/or spinach lines SMB66-1144F and SMBS012-1188Mmay be defined as forming at least about 97% of the total seed,including at least about 98%, 99% or more of the seed. The seedpopulation may be separately grown to provide an essentially homogeneouspopulation of spinach plants designated SV8545VC and/or spinach linesSMB66-1144F and SMBS012-1188M.

In yet another aspect of the invention, a tissue culture of regenerablecells of a spinach plant of hybrid SV8545VC and/or spinach linesSMB66-1144F and SMBS012-1188M is provided. The tissue culture willpreferably be capable of regenerating spinach plants capable ofexpressing all of the physiological and morphological characteristics ofthe starting plant, and of regenerating plants having substantially thesame genotype as the starting plant. Examples of some of thephysiological and morphological characteristics of the hybrid SV8545VCand/or spinach lines SMB66-1144F and SMBS012-1188M include those traitsset forth in the tables herein. The regenerable cells in such tissuecultures may be derived, for example, from embryos, meristems,cotyledons, pollen, leaves, anthers, roots, root tips, pistils, flowers,seed and stalks. Still further, the present invention provides spinachplants regenerated from a tissue culture of the invention, the plantshaving all the physiological and morphological characteristics of hybridSV8545VC and/or spinach lines SMB66-1144F and SMBS012-1188M.

In still yet another aspect of the invention, processes are provided forproducing spinach seeds, plants and fruit, which processes generallycomprise crossing a first parent spinach plant with a second parentspinach plant, wherein at least one of the first or second parentspinach plants is a plant of spinach line SMB66-1144F or SMBS012-1188M.These processes may be further exemplified as processes for preparinghybrid spinach seed or plants, wherein a first spinach plant is crossedwith a second spinach plant of a different, distinct genotype to providea hybrid that has, as one of its parents, a plant of spinach lineSMB66-1144F or SMBS012-1188M. In these processes, crossing will resultin the production of seed. The seed production occurs regardless ofwhether the seed is collected or not.

In one embodiment of the invention, the first step in “crossing”comprises planting seeds of a first and second parent spinach plant,often in proximity so that pollination will occur for example, mediatedby insect vectors. Alternatively, pollen can be transferred manually.Where the plant is self-pollinated, pollination may occur without theneed for direct human intervention other than plant cultivation.

A second step may comprise cultivating or growing the seeds of first andsecond parent spinach plants into plants that bear flowers. A third stepmay comprise preventing self-pollination of the plants, such as byemasculating the flowers (i.e., killing or removing the pollen).

A fourth step for a hybrid cross may comprise cross-pollination betweenthe first and second parent spinach plants. Yet another step comprisesharvesting the seeds from at least one of the parent spinach plants. Theharvested seed can be grown to produce a spinach plant or hybrid spinachplant.

The present invention also provides the spinach seeds and plantsproduced by a process that comprises crossing a first parent spinachplant with a second parent spinach plant, wherein at least one of thefirst or second parent spinach plants is a plant of spinach hybridSV8545VC and/or spinach lines SMB66-1144F and SMBS012-1188M. In oneembodiment of the invention, spinach seed and plants produced by theprocess are first generation (F₁) hybrid spinach seed and plantsproduced by crossing a plant in accordance with the invention withanother, distinct plant. The present invention further contemplatesplant parts of such an F₁ hybrid spinach plant, and methods of usethereof. Therefore, certain exemplary embodiments of the inventionprovide an F₁ hybrid spinach plant and seed thereof.

In still yet another aspect, the present invention provides a method ofproducing a plant derived from hybrid SV8545VC and/or spinach linesSMB66-1144F and SMBS012-1188M, the method comprising the steps of: (a)preparing a progeny plant derived from hybrid SV8545VC and/or spinachlines SMB66-1144F and SMBS012-1188M, wherein said preparing comprisescrossing a plant of the hybrid SV8545VC and/or spinach lines SMB66-1144Fand SMBS012-1188M with a second plant; and (b) crossing the progenyplant with itself or a second plant to produce a seed of a progeny plantof a subsequent generation. In further embodiments, the method mayadditionally comprise: (c) growing a progeny plant of a subsequentgeneration from said seed of a progeny plant of a subsequent generationand crossing the progeny plant of a subsequent generation with itself ora second plant; and repeating the steps for an additional 3-10generations to produce a plant derived from hybrid SV8545VC and/orspinach lines SMB66-1144F and SMBS012-1188M. The plant derived fromhybrid SV8545VC and/or spinach lines SMB66-1144F and SMBS012-1188M maybe an inbred line, and the aforementioned repeated crossing steps may bedefined as comprising sufficient inbreeding to produce the inbred line.In the method, it may be desirable to select particular plants resultingfrom step (c) for continued crossing according to steps (b) and (c). Byselecting plants having one or more desirable traits, a plant derivedfrom hybrid SV8545VC and/or spinach lines SMB66-1144F and SMBS012-1188Mis obtained which possesses some of the desirable traits of theline/hybrid as well as potentially other selected traits.

In certain embodiments, the present invention provides a method ofproducing food or feed comprising: (a) obtaining a plant of spinachhybrid SV8545VC and/or spinach lines SMB66-1144F and SMBS012-1188M,wherein the plant has been cultivated to maturity, and (b) collecting atleast one spinach from the plant.

In still yet another aspect of the invention, the genetic complement ofspinach hybrid SV8545VC and/or spinach lines SMB66-1144F andSMBS012-1188M is provided. The phrase “genetic complement” is used torefer to the aggregate of nucleotide sequences, the expression of whichsequences defines the phenotype of, in the present case, a spinachplant, or a cell or tissue of that plant. A genetic complement thusrepresents the genetic makeup of a cell, tissue or plant, and a hybridgenetic complement represents the genetic make up of a hybrid cell,tissue or plant. The invention thus provides spinach plant cells thathave a genetic complement in accordance with the spinach plant cellsdisclosed herein, and seeds and plants containing such cells.

Plant genetic complements may be assessed by genetic marker profiles,and by the expression of phenotypic traits that are characteristic ofthe expression of the genetic complement, e.g., isozyme typing profiles.It is understood that hybrid SV8545VC and/or spinach lines SMB66-1144Fand SMBS012-1188M could be identified by any of the many well knowntechniques such as, for example, Simple Sequence Length Polymorphisms(SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990),Randomly Amplified Polymorphic DNAs (RAPDs), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified FragmentLength Polymorphisms (AFLPs) (EP 534 858, specifically incorporatedherein by reference in its entirety), and Single NucleotidePolymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).

In still yet another aspect, the present invention provides hybridgenetic complements, as represented by spinach plant cells, tissues,plants, and seeds, formed by the combination of a haploid geneticcomplement of a spinach plant of the invention with a haploid geneticcomplement of a second spinach plant, preferably, another, distinctspinach plant. In another aspect, the present invention provides aspinach plant regenerated from a tissue culture that comprises a hybridgenetic complement of this invention.

Any embodiment discussed herein with respect to one aspect of theinvention applies to other aspects of the invention as well, unlessspecifically noted.

The term “about” is used to indicate that a value includes the standarddeviation of the mean for the device or method being employed todetermine the value. The use of the term “or” in the claims is used tomean “and/or” unless explicitly indicated to refer to alternatives onlyor the alternatives are mutually exclusive. When used in conjunctionwith the word “comprising” or other open language in the claims, thewords “a” and “an” denote “one or more,” unless specifically notedotherwise. The terms “comprise,” “have” and “include” are open-endedlinking verbs. Any forms or tenses of one or more of these verbs, suchas “comprises,” “comprising,” “has,” “having,” “includes” and“including,” are also open-ended. For example, any method that“comprises,” “has” or “includes” one or more steps is not limited topossessing only those one or more steps and also covers other unlistedsteps. Similarly, any plant that “comprises,” “has” or “includes” one ormore traits is not limited to possessing only those one or more traitsand covers other unlisted traits.

Other objects, features and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and any specificexamples provided, while indicating specific embodiments of theinvention, are given by way of illustration only, since various changesand modifications within the spirit and scope of the invention willbecome apparent to those skilled in the art from this detaileddescription.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants,seeds and derivatives of spinach hybrid SV8545VC, spinach lineSMB66-1144F or SMBS012-1188M.

Spinach (Spinacia oleracea L.) hybrid SV8545VC is a mid early bolting,smooth leaf type variety with spineless seeds and resistance againstPeronospora farinosa f.sp. spinaciae races Pfs 1 to Pfs 13 and 15.Furthermore the hybrid is phenotypically distinct from well-knownmaterial by a combination of Pfs 1 to Pfs 13 and 15 resistances andleaves which stay green for a long period.

Hybrid SV8545VC is believed to most closely resemble Seminis varietyTonga. The comparative characteristics that mostly distinguish to twohybrids include, but may not be limited to, are resistances, maturity(growth speed) and curving of the margin. Hybrid SV8545VC has adifferent resistance pattern, maturity and curving of the margincompared to Tonga.

The development of the line can be summarized as follows.

A. ORIGIN AND BREEDING HISTORY OF SPINACH HYBRID SV8545VC

Hybrid SV8545VC was developed in Year 1 by crossing female lineSMB66-1144F with male line SMBS012-1188M. The final outcome was an earlybolting hybrid with smooth leaves with resistances against downy mildewraces Pfs 1 to Pfs 13 and 15. Observation during the variety selectionsin Year 1 until Year 5 confirmed that SV845VC is uniform and stable. Asis true with other spinach varieties, a small percentage of off-typescan occur for almost any characteristics during the course of repeatedmultiplications. However, no variants were observed for SV8545VC.

The parent lines are uniform and stable, as is a hybrid producedtherefrom. A small percentage of variants can occur within commerciallyacceptable limits for almost any characteristic during the course ofrepeated multiplication. However no variants are expected.

B. PHYSIOLOGICAL AND MORPHOLOGICAL CHARACTERISTICS OF SPINACH HYBRIDSV8545VC, AND SPINACH LINES SMB66-1144F AND SMBS012-1188M

In accordance with one aspect of the present invention, there isprovided a plant having the physiological and morphologicalcharacteristics of spinach hybrid SV8545VC and the parent lines thereof.A description of the physiological and morphological characteristics ofsuch plants is presented in Tables 1-3.

TABLE 1 Physiological and Morphological Characteristics of HybridSV8545VC. CHARACTERISTIC SV8545VC Tonga Ploidy diploid diploid SeedlingCotyledon Length of cotyledon medium medium Width (mm)  9.6 mm  8.1 mmLength (mm) 77.5 mm 73.4 mm Tip pointed pointed Color medium greenmedium green Color Chart Name RHS Color Chart Value 146B 146B Leaf(First Foliage Leaves) Shape ovate ovate Base V-shape V-shape Marginflat flat Upper Surface Color dark green (Long dark green (Long StandingBloomsdale) Standing Bloomsdale) Color Chart Name RHS Color Chart Value146A 146A Lower Surface Color lighter lighter (Compared with uppersurface) Color Chart Name RHS Color Chart Value 146C 146B MaturityGrowth Rate slow (Norgreen) slow (Norgreen) Days from planting to 42 46prime market stage Plant (Prime Market Stage) Habit semi-erect (Longsemi-erect (Long Standing Bloomsdale) Standing Bloomsdale) Size large(Giant Nobel) large (Giant Nobel) Spread (cm) 34.6 cm 34.1 cm Height(cm) 14.6 cm 12.9 cm Leaf (Prime Market Stage) Surface smooth (Viroflay)smooth (Viroflay) Anthocyanin coloration of absent absent petioles andveins Blade blistering medium medium Blade lobing weak weak Petiole:attitude semi-erect semi-erect Blade attitude semi-erect semi-erect Leafshape ovate ovate Leaf Blade: shape Medium ovate Medium ovate (excludingbasal lobes) (prime market stage) Blade curving of margin flat flatBlade shape of apex rounded rounded Blade shape in longitudinal convexconvex section Base lobed lobed Margin flat flat Upper Surface Colormedium green medium green Color Chart Name RHS Color Chart Value 146A  146A+ Lower Surface Color lighter lighter (Compared with uppersurface) Color Chart Name RHS Color Chart Value 146B 146B Luster dulldull Blade size medium medium Petiole color light green light greenPetiole color RHS 144A 144A Petiole red pigmentation absent absent cmpetiole length to the 13.0 cm 12.4 cm blade mm petiole diameter  7.0 cm 7.9 cm Petiole diameter medium large Seed Stalk Development Start ofBolting (10% of medium medium plants) Height of Stalk (cm) 82.3 cm 82.8cm Plants that are Female  0-10% [Monnopa]  0-10% [Monnopa] Proportionof female plants absent or very low absent or very low Plants that areMale 0-10% [Monnopa, 0-10% [Monnopa, Parrot] Parrot] Proportion of maleplants absent or very low absent or very low Plants that are Monoecious91-100% [Monnopa] 91-100% [Monnopa] Proportion of monoecious very highvery high plants Seed Surface smooth smooth Spines (harvested seed)absent [Resistoflay] absent [Resistoflay] These are typical values.Values may vary due to environment. Other values that are substantiallyequivalent are also within the scope of the invention.

TABLE 2 Physiological and Morphological Characteristics of LineSMB66-1144F. CHARACTERISTIC SMB-66-1144F SSB-66-1087F Ploidy diploiddiploid Seedling Cotyledon Length of cotyledon medium medium Width (mm) 5.3 mm  5.6 mm Length (mm) 47.7 mm 47.7 mm Tip pointed pointed Colormedium green medium green Color Chart Name RHS RHS Color Chart Value  146b+   146b+ Leaf (First Foliage Leaves) Shape ovate ovate BaseV-shape V-shape Tip round round-pointed Margin flat flat Upper SurfaceColor medium green medium green (Giant Nobel) (Giant Nobel) Color ChartName RHS RHS Color Chart Value 146a 146a Lower Surface Color lighterlighter (Compared with upper surface) Color Chart Name RHS RHS ColorChart Value 146b 146b Maturity Growth Rate slow (Norgreen) medium (LongStanding Bloomsdale) Days from planting to prime 46 54 market stagePlant (Prime Market Stage) Habit flat (Viroflay) semi-erect (LongStanding Bloomsdale) Size medium medium Spread (cm) 42.5 cm 44.9 cmHeight (cm) 17.9 cm 13.9 cm Leaf (Prime Market Stage) Surface smooth(Viroflay) smooth (Viroflay) Anthocyanin coloration of absent absentpetioles and veins Blade intensity of green color dark dark Bladeblistering weak weak Blade lobing weak absent or very weak Petiole:attitude semi-erect semi-erect Petiole: length (prime market mediummedium stage) Blade attitude semi-pendulous horizontal Leaf shape ovateovate Blade shape (excluding basal medium ovate medium ovate lobes)Blade curving of margin flat recurved Blade shape of apex obtuse roundedBlade shape in longitudinal convex flat section Base straight straightTip round-pointed round Upper Surface Color medium green medium greenColor Chart Name RHS RHS Color Chart Value 146a 146a+ Lower SurfaceColor lighter lighter (Compared with upper surface) Color Chart Name RHSRHS Color Chart Value 146b 146b Luster dull dull Blade size mediummedium Petiole color light green light green Petiole color RHS 144a 144aPetiole red pigmentation absent absent cm petiole length to the blade11.2 cm 11.5 cm mm petiole diameter  5.9 mm  6.8 mm Petiole diametermedium medium Seed Stalk Development Start of Bolting (10% of mediumlate plants) Time of start of bolting (for medium late spring sown crop,15% of plants) Height of Stalk (cm) 34.2 cm 43.5 cm Leaves on Stalk ofFemale few or none few or none Plant Plants that are Female 0-10% 0-10%Proportion of Female Plants absent or very absent or very low low Plantsthat are Male 0-10% 0-10% Proportion of Male Plants absent or veryabsent or very low low Plants that are Monoecious 91-100% 91-100%Proportion of Monoecious very high very high Plants Seed Surface smoothsmooth Spines (harvested seed) absent absent [Resistoflay] [Resistoflay]These are typical values. Values may vary due to environment. Othervalues that are substantially equivalent are also within the scope ofthe invention.

TABLE 3 Physiological and Morphological Characteristics of SMBS012-1188MCHARACTERISTIC SMBS012-1188M SMB66-0027F Ploidy diploid diploid SeedlingCotyledon Length of cotyledon medium medium Width (mm)  9.3 mm  9.1 mmLength (mm) 87.8 mm 81.4 mm Tip pointed pointed Color medium greenmedium green/light green Color Chart Name RHS RHS Color Chart Value146bc 146bc Leaf (First Foliage Leaves) Shape ovate ovate Base straightstraight Tip round-pointed round Margin flat flat Upper Surface Colormedium green medium green (Giant Nobel) (Giant Nobel) Color Chart NameRHS RHS Color Chart Value 146a 146b Lower Surface Color lighter lighter(Compared with upper surface) Color Chart Name RHS RHS Color Chart Value146b 146c Maturity Growth Rate fast (Dixie Market) fast (Dixie Market)Days from planting to prime 41 35 market stage Plant (Prime MarketStage) Habit semi-erect (Long semi-erect (Long Standing StandingBloomsdale) Bloomsdale) Size medium medium Spread (cm) 30.8 cm 31.0 cmHeight (cm) 11.7 cm  8.8 cm Leaf (Prime Market Stage) Surface smooth(Viroflay) smooth (Viroflay) Anthocyanin coloration of absent absentpetioles and veins Blade intensity of green color medium medium Bladelobing weak weak Petiole: attitude semi-erect semi-erect Petiole: length(prime market medium medium stage) Blade attitude horizontalsemi-pendulous Leaf shape ovate ovate Blade shape (excluding basal broadovate medium ovate lobes) Blade curving of margin flat flat Blade shapeof apex obtuse obtuse Blade shape in longitudinal flat convex sectionBase lobed lobed Tip round-pointed round-pointed Margin slightly curvedflat Upper Surface Color medium green medium green Color Chart Name RHSRHS Color Chart Value 147a 147a Lower Surface Color lighter lighter(Compared with upper surface) Color Chart Name RHS RHS Color Chart Value146B 146b Luster dull dull Blade size medium medium Petiole color lightgreen light green Petiole color RHS 144A 144a Petiole red pigmentationabsent absent cm petiole length to the blade 10.3 cm 8.1 cm mm petiolediameter 6.5 mm 6.4 mm Petiole diameter medium medium Seed StalkDevelopment Start of Bolting (10% of medium medium plants) Time of startof bolting (for medium medium spring sown crop, 15% of plants) Height ofStalk (cm) 75.8 cm 66.6 cm Leaves on Stalk of Male few or none few ornone Plant Plants that are Female 0-10% 0-10% Proportion of FemalePlants absent or very low absent or very low Plants that are Male 0-10%0-10% Proportion of Male Plants absent or very low absent or very lowPlants that are Monoecious 91-100% 91-100% Proportion of Monoecious veryhigh very high Plants Seed Surface smooth smooth Spines (harvested seed)absent [Resistoflay] absent [Resistoflay] These are typical values.Values may vary due to environment. Other values that are substantiallyequivalent are also within the scope of the invention.

C. BREEDING SPINACH PLANTS

One aspect of the current invention concerns methods for producing seedof spinach hybrid SV8545VC involving crossing spinach lines SMB66-1144Fand SMBS012-1188M. Alternatively, in other embodiments of the invention,hybrid SV8545VC, line SMB66-1144F or line SMBS012-1188M may be crossedwith itself or with any second plant. Such methods can be used forpropagation of hybrid SV8545VC and/or the spinach lines SMB66-1144F andSMBS012-1188M, or can be used to produce plants that are derived fromhybrid SV8545VC and/or the spinach lines SMB66-1144F and SMBS012-1188M.Plants derived from hybrid SV8545VC and/or the spinach lines SMB66-1144Fand SMBS012-1188M may be used, in certain embodiments, for thedevelopment of new spinach varieties.

The development of new varieties using one or more starting varieties iswell known in the art. In accordance with the invention, novel varietiesmay be created by crossing hybrid SV8545VC followed by multiplegenerations of breeding according to such well known methods. Newvarieties may be created by crossing with any second plant. In selectingsuch a second plant to cross for the purpose of developing novel lines,it may be desired to choose those plants which either themselves exhibitone or more selected desirable characteristics or which exhibit thedesired characteristic(s) when in hybrid combination. Once initialcrosses have been made, inbreeding and selection take place to producenew varieties. For development of a uniform line, often five or moregenerations of selfing and selection are involved.

Uniform lines of new varieties may also be developed by way ofdouble-haploids. This technique allows the creation of true breedinglines without the need for multiple generations of selfing andselection. In this manner true breeding lines can be produced in aslittle as one generation. Haploid embryos may be produced frommicrospores, pollen, anther cultures, or ovary cultures. The haploidembryos may then be doubled autonomously, or by chemical treatments(e.g. colchicine treatment). Alternatively, haploid embryos may be growninto haploid plants and treated to induce chromosome doubling. In eithercase, fertile homozygous plants are obtained. In accordance with theinvention, any of such techniques may be used in connection with a plantof the invention and progeny thereof to achieve a homozygous line.

Backcrossing can also be used to improve an inbred plant. Backcrossingtransfers a specific desirable trait from one inbred or non-inbredsource to an inbred that lacks that trait. This can be accomplished, forexample, by first crossing a superior inbred (A) (recurrent parent) to adonor inbred (non-recurrent parent), which carries the appropriate locusor loci for the trait in question. The progeny of this cross are thenmated back to the superior recurrent parent (A) followed by selection inthe resultant progeny for the desired trait to be transferred from thenon-recurrent parent. After five or more backcross generations withselection for the desired trait, the progeny have the characteristicbeing transferred, but are like the superior parent for most or almostall other loci. The last backcross generation would be selfed to givepure breeding progeny for the trait being transferred.

The plants of the present invention are particularly well suited for thedevelopment of new lines based on the elite nature of the geneticbackground of the plants. In selecting a second plant to cross withSV8545VC and/or spinach lines SMB66-1144F and SMBS012-1188M for thepurpose of developing novel spinach lines, it will typically bepreferred to choose those plants which either themselves exhibit one ormore selected desirable characteristics or which exhibit the desiredcharacteristic(s) when in hybrid combination. Examples of desirabletraits may include, in specific embodiments, high seed yield, high seedgermination, seedling vigor, high fruit yield, disease tolerance orresistance, and adaptability for soil and climate conditions.Consumer-driven traits, such as a fruit shape, color, texture, and tasteare other examples of traits that may be incorporated into new lines ofspinach plants developed by this invention.

D. PERFORMANCE CHARACTERISTICS

As described above, hybrid SV8545VC exhibits desirable traits, asconferred by spinach lines SMB66-1144F and SMBS012-1188M. Theperformance characteristics of hybrid SV8545VC and spinach linesSMB66-1144F and SMBS012-1188M were the subject of an objective analysisof the performance traits relative to other varieties. The results ofthe analysis are presented below.

Parent line SMB66-1144F can be characterized as an early bolting linethat has spineless seeds (smooth) and with a resistance to downy mildew(Peronospora farinosa f.sp. spinaciae (Pfs) races Pfs 1 to Pfs 10.Parental line SMB66-1144F is believed to most closely resemble Monsantoparental line SSB-66-1087F. The comparative characteristics that mostlydistinguish to two lines include, but may not be limited to Maturity,Harvest Window and Production. The candidate line SMB66-1144F has afaster maturity, better harvest window and more production compared toSMB-66-1087F.

Parent line SMBS012-1188M can be characterized as an early bolting linethat has spineless seeds (smooth) and with a resistance to downy mildew(Peronospora farinosa f.sp. spinaciae (Pfs) races Pfs 1 to Pfs 13.Furthermore, the line is phenotypically distinct from all well-knownmaterial by a combination of high productivity (big leaves), and longstems. Parental line SMBS012-1188M is believed to most closely resembleMonsanto parental line SMB66-0027F. The comparative characteristics thatmostly distinguish to two lines include, but may not be limited to, sexexpression, resistances and productivity. The candidate lineSMBS012-1188M flowers with mostly male flowers, is resistant to downymildew races pfs1-13 and is more productive compared to SMB66-0027F.

TABLE 4 Comparison of SV8545VC with Tonga SV8545VC Tonga Resistances Pfs1-13, 15 Pfs 1-11, 13, 15 Maturity Earlier Later Curving of the marginRecurved Flat Location: Wageningen, the Netherlands 2010 until 2014.

TABLE 5 Comparison SMB66-1144F with SSB-66-1087F Maturity Harvest WindowProduction SSB66-1144F Fastest Biggest Most SSB66-1087F Slowest SmallestLeast Location: Wageningen, the Netherlands.

TABLE 6 Comparison SMBS012-1188M with SMB66-0027F Sex ExpressionResistances Production SMBS012-1188M Mostly male flowers Pfs 1-13 Mostproductive SMB66-0027F Mostly female flowers Pfs 1-4  Least productiveLocation: Wageningen, the Netherlands.

E. FURTHER EMBODIMENTS OF THE INVENTION

In certain aspects of the invention, plants described herein areprovided modified to include at least a first desired heritable trait.Such plants may, in one embodiment, be developed by a plant breedingtechnique called backcrossing, wherein essentially all of themorphological and physiological characteristics of a variety arerecovered in addition to a genetic locus transferred into the plant viathe backcrossing technique. The term single locus converted plant asused herein refers to those spinach plants which are developed by aplant breeding technique called backcrossing, wherein essentially all ofthe morphological and physiological characteristics of a variety arerecovered in addition to the single locus transferred into the varietyvia the backcrossing technique. By essentially all of the morphologicaland physiological characteristics, it is meant that the characteristicsof a plant are recovered that are otherwise present when compared in thesame environment, other than an occasional variant trait that mightarise during backcrossing or direct introduction of a transgene.

Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the present variety. The parentalspinach plant which contributes the locus for the desired characteristicis termed the nonrecurrent or donor parent. This terminology refers tothe fact that the nonrecurrent parent is used one time in the backcrossprotocol and therefore does not recur. The parental spinach plant towhich the locus or loci from the nonrecurrent parent are transferred isknown as the recurrent parent as it is used for several rounds in thebackcrossing protocol.

In a typical backcross protocol, the original variety of interest(recurrent parent) is crossed to a second variety (nonrecurrent parent)that carries the single locus of interest to be transferred. Theresulting progeny from this cross are then crossed again to therecurrent parent and the process is repeated until a spinach plant isobtained wherein essentially all of the morphological and physiologicalcharacteristics of the recurrent parent are recovered in the convertedplant, in addition to the single transferred locus from the nonrecurrentparent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single locus of the recurrent variety ismodified or substituted with the desired locus from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphologicalconstitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross; one ofthe major purposes is to add some commercially desirable trait to theplant. The exact backcrossing protocol will depend on the characteristicor trait being altered and the genetic distance between the recurrentand nonrecurrent parents. Although backcrossing methods are simplifiedwhen the characteristic being transferred is a dominant allele, arecessive allele, or an additive allele (between recessive anddominant), may also be transferred. In this instance it may be necessaryto introduce a test of the progeny to determine if the desiredcharacteristic has been successfully transferred.

In one embodiment, progeny spinach plants of a backcross in which aplant described herein is the recurrent parent comprise (i) the desiredtrait from the non-recurrent parent and (ii) all of the physiologicaland morphological characteristics of spinach the recurrent parent asdetermined at the 5% significance level when grown in the sameenvironmental conditions.

New varieties can also be developed from more than two parents. Thetechnique, known as modified backcrossing, uses different recurrentparents during the backcrossing. Modified backcrossing may be used toreplace the original recurrent parent with a variety having certain moredesirable characteristics or multiple parents may be used to obtaindifferent desirable characteristics from each.

With the development of molecular markers associated with particulartraits, it is possible to add additional traits into an established germline, such as represented here, with the end result being substantiallythe same base germplasm with the addition of a new trait or traits.Molecular breeding, as described in Moose and Mumm, 2008 (PlantPhysiology, 147: 969-977), for example, and elsewhere, provides amechanism for integrating single or multiple traits or QTL into an eliteline. This molecular breeding-facilitated movement of a trait or traitsinto an elite line may encompass incorporation of a particular genomicfragment associated with a particular trait of interest into the eliteline by the mechanism of identification of the integrated genomicfragment with the use of flanking or associated marker assays. In theembodiment represented here, one, two, three or four genomic loci, forexample, may be integrated into an elite line via this methodology. Whenthis elite line containing the additional loci is further crossed withanother parental elite line to produce hybrid offspring, it is possibleto then incorporate at least eight separate additional loci into thehybrid. These additional loci may confer, for example, such traits as adisease resistance or a fruit quality trait. In one embodiment, eachlocus may confer a separate trait. In another embodiment, loci may needto be homozygous and exist in each parent line to confer a trait in thehybrid. In yet another embodiment, multiple loci may be combined toconfer a single robust phenotype of a desired trait.

Many single locus traits have been identified that are not regularlyselected for in the development of a new inbred but that can be improvedby backcrossing techniques. Single locus traits may or may not betransgenic; examples of these traits include, but are not limited to,herbicide resistance, resistance to bacterial, fungal, or viral disease,insect resistance, modified fatty acid or carbohydrate metabolism, andaltered nutritional quality. These comprise genes generally inheritedthrough the nucleus.

Direct selection may be applied where the single locus acts as adominant trait. For this selection process, the progeny of the initialcross are assayed for viral resistance and/or the presence of thecorresponding gene prior to the backcrossing. Selection eliminates anyplants that do not have the desired gene and resistance trait, and onlythose plants that have the trait are used in the subsequent backcross.This process is then repeated for all additional backcross generations.

Selection of spinach plants for breeding is not necessarily dependent onthe phenotype of a plant and instead can be based on geneticinvestigations. For example, one can utilize a suitable genetic markerwhich is closely genetically linked to a trait of interest. One of thesemarkers can be used to identify the presence or absence of a trait inthe offspring of a particular cross, and can be used in selection ofprogeny for continued breeding. This technique is commonly referred toas marker assisted selection. Any other type of genetic marker or otherassay which is able to identify the relative presence or absence of atrait of interest in a plant can also be useful for breeding purposes.Procedures for marker assisted selection are well known in the art. Suchmethods will be of particular utility in the case of recessive traitsand variable phenotypes, or where conventional assays may be moreexpensive, time consuming or otherwise disadvantageous. Types of geneticmarkers which could be used in accordance with the invention include,but are not necessarily limited to, Simple Sequence Length Polymorphisms(SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990),Randomly Amplified Polymorphic DNAs (RAPDs), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified FragmentLength Polymorphisms (AFLPs) (EP 534 858, specifically incorporatedherein by reference in its entirety), and Single NucleotidePolymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).

F. PLANTS DERIVED BY GENETIC ENGINEERING

Many useful traits that can be introduced by backcrossing, as well asdirectly into a plant, are those which are introduced by genetictransformation techniques. Genetic transformation may therefore be usedto insert a selected transgene into a plant of the invention or may,alternatively, be used for the preparation of transgenes which can beintroduced by backcrossing. Methods for the transformation of plantsthat are well known to those of skill in the art and applicable to manycrop species include, but are not limited to, electroporation,microprojectile bombardment, Agrobacterium-mediated transformation anddirect DNA uptake by protoplasts.

To effect transformation by electroporation, one may employ eitherfriable tissues, such as a suspension culture of cells or embryogeniccallus or alternatively one may transform immature embryos or otherorganized tissue directly. In this technique, one would partiallydegrade the cell walls of the chosen cells by exposing them topectin-degrading enzymes (pectolyases) or mechanically wound tissues ina controlled manner.

An efficient method for delivering transforming DNA segments to plantcells is microprojectile bombardment. In this method, particles arecoated with nucleic acids and delivered into cells by a propellingforce. Exemplary particles include those comprised of tungsten,platinum, and preferably, gold. For the bombardment, cells in suspensionare concentrated on filters or solid culture medium. Alternatively,immature embryos or other target cells may be arranged on solid culturemedium. The cells to be bombarded are positioned at an appropriatedistance below the macroprojectile stopping plate.

An illustrative embodiment of a method for delivering DNA into plantcells by acceleration is the Biolistics Particle Delivery System, whichcan be used to propel particles coated with DNA or cells through ascreen, such as a stainless steel or Nytex screen, onto a surfacecovered with target cells. The screen disperses the particles so thatthey are not delivered to the recipient cells in large aggregates.Microprojectile bombardment techniques are widely applicable, and may beused to transform virtually any plant species.

Agrobacterium-mediated transfer is another widely applicable system forintroducing gene loci into plant cells. An advantage of the technique isthat DNA can be introduced into whole plant tissues, thereby bypassingthe need for regeneration of an intact plant from a protoplast. ModernAgrobacterium transformation vectors are capable of replication in E.coli as well as Agrobacterium, allowing for convenient manipulations(Klee et al., Bio-Technology, 3(7):637-642, 1985). Moreover, recenttechnological advances in vectors for Agrobacterium-mediated genetransfer have improved the arrangement of genes and restriction sites inthe vectors to facilitate the construction of vectors capable ofexpressing various polypeptide coding genes. The vectors described haveconvenient multi-linker regions flanked by a promoter and apolyadenylation site for direct expression of inserted polypeptidecoding genes. Additionally, Agrobacterium containing both armed anddisarmed Ti genes can be used for transformation.

In those plant strains where Agrobacterium-mediated transformation isefficient, it is the method of choice because of the facile and definednature of the gene locus transfer. The use of Agrobacterium-mediatedplant integrating vectors to introduce DNA into plant cells is wellknown in the art (Fraley et al., Bio/Technology, 3:629-635, 1985; U.S.Pat. No. 5,563,055).

Transformation of plant protoplasts also can be achieved using methodsbased on calcium phosphate precipitation, polyethylene glycol treatment,electroporation, and combinations of these treatments (see, e.g.,Potrykus et al., Mol. Gen. Genet., 199:183-188, 1985; Omirulleh et al.,Plant Mol. Biol., 21(3):415-428, 1993; Fromm et al., Nature,312:791-793, 1986; Uchimiya et al., Mol. Gen. Genet., 204:204, 1986;Marcotte et al., Nature, 335:454, 1988). Transformation of plants andexpression of foreign genetic elements is exemplified in Choi et al.(Plant Cell Rep., 13: 344-348, 1994), and Ellul et al. (Theor. Appl.Genet., 107:462-469, 2003).

A number of promoters have utility for plant gene expression for anygene of interest including but not limited to selectable markers,scoreable markers, genes for pest tolerance, disease resistance,nutritional enhancements and any other gene of agronomic interest.Examples of constitutive promoters useful for plant gene expressioninclude, but are not limited to, the cauliflower mosaic virus (CaMV)P-35S promoter, which confers constitutive, high-level expression inmost plant tissues (see, e.g., Odel et al., Nature, 313:810, 1985),including in monocots (see, e.g., Dekeyser et al., Plant Cell, 2:591,1990; Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990); a tandemlyduplicated version of the CaMV 35S promoter, the enhanced 35S promoter(P-e35S); 1 the nopaline synthase promoter (An et al., Plant Physiol.,88:547, 1988); the octopine synthase promoter (Fromm et al., Plant Cell,1:977, 1989); and the figwort mosaic virus (P-FMV) promoter as describedin U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter(P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem;the cauliflower mosaic virus 19S promoter; a sugarcane bacilliform viruspromoter; a commelina yellow mottle virus promoter; and other plant DNAvirus promoters known to express in plant cells.

A variety of plant gene promoters that are regulated in response toenvironmental, hormonal, chemical, and/or developmental signals can alsobe used for expression of an operably linked gene in plant cells,including promoters regulated by (1) heat (Callis et al., PlantPhysiol., 88:965, 1988), (2) light (e.g., pea rbcS-3A promoter,Kuhlemeier et al., Plant Cell, 1:471, 1989; maize rbcS promoter,Schaffner and Sheen, Plant Cell, 3:997, 1991; or chlorophyll a/b-bindingprotein promoter, Simpson et al., EMBO J., 4:2723, 1985), (3) hormones,such as abscisic acid (Marcotte et al., Plant Cell, 1:969, 1989), (4)wounding (e.g., wunl, Siebertz et al., Plant Cell, 1:961, 1989); or (5)chemicals such as methyl jasmonate, salicylic acid, or Safener. It mayalso be advantageous to employ organ-specific promoters (e.g., Roshal etal., EMBO J., 6:1155, 1987; Schernthaner et al., EMBO J., 7:1249, 1988;Bustos et al., Plant Cell, 1:839, 1989).

Exemplary nucleic acids which may be introduced to plants of thisinvention include, for example, DNA sequences or genes from anotherspecies, or even genes or sequences which originate with or are presentin the same species, but are incorporated into recipient cells bygenetic engineering methods rather than classical reproduction orbreeding techniques. However, the term “exogenous” is also intended torefer to genes that are not normally present in the cell beingtransformed, or perhaps simply not present in the form, structure, etc.,as found in the transforming DNA segment or gene, or genes which arenormally present and that one desires to express in a manner thatdiffers from the natural expression pattern, e.g., to over-express.Thus, the term “exogenous” gene or DNA is intended to refer to any geneor DNA segment that is introduced into a recipient cell, regardless ofwhether a similar gene may already be present in such a cell. The typeof DNA included in the exogenous DNA can include DNA which is alreadypresent in the plant cell, DNA from another plant, DNA from a differentorganism, or a DNA generated externally, such as a DNA sequencecontaining an antisense message of a gene, or a DNA sequence encoding asynthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and couldpotentially be introduced into a spinach plant according to theinvention. Non-limiting examples of particular genes and correspondingphenotypes one may choose to introduce into a spinach plant include oneor more genes for insect tolerance, such as a Bacillus thuringiensis(B.t.) gene, pest tolerance such as genes for fungal disease control,herbicide tolerance such as genes conferring glyphosate tolerance, andgenes for quality improvements such as yield, nutritional enhancements,environmental or stress tolerances, or any desirable changes in plantphysiology, growth, development, morphology or plant product(s). Forexample, structural genes would include any gene that confers insecttolerance including but not limited to a Bacillus insect control proteingene as described in WO 99/31248, herein incorporated by reference inits entirety, U.S. Pat. No. 5,689,052, herein incorporated by referencein its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, hereinincorporated by reference in their entirety. In another embodiment, thestructural gene can confer tolerance to the herbicide glyphosate asconferred by genes including, but not limited to Agrobacterium strainCP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat.No. 5,633,435, herein incorporated by reference in its entirety, orglyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No.5,463,175, herein incorporated by reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes byencoding a non-translatable RNA molecule that causes the targetedinhibition of expression of an endogenous gene, for example viaantisense- or cosuppression-mediated mechanisms (see, for example, Birdet al., Biotech. Gen. Engin. Rev., 9:207, 1991). The RNA could also be acatalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desiredendogenous mRNA product (see for example, Gibson and Shillito, Mol.Biotech., 7:125, 1997). Thus, any gene which produces a protein or mRNAwhich expresses a phenotype or morphology change of interest is usefulfor the practice of the present invention.

G. DEFINITIONS

In the description and tables herein, a number of terms are used. Inorder to provide a clear and consistent understanding of thespecification and claims, the following definitions are provided:

Allele: Any of one or more alternative forms of a gene locus, all ofwhich alleles relate to one trait or characteristic. In a diploid cellor organism, the two alleles of a given gene occupy corresponding locion a pair of homologous chromosomes.

Backcrossing: A process in which a breeder repeatedly crosses hybridprogeny, for example a first generation hybrid (F₁), back to one of theparents of the hybrid progeny. Backcrossing can be used to introduce oneor more single locus conversions from one genetic background intoanother.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes fromdifferent plants.

Diploid: A cell or organism having two sets of chromosomes.

Emasculate: The removal of plant male sex organs or the inactivation ofthe organs with a cytoplasmic or nuclear genetic factor or a chemicalagent conferring male sterility.

Enzymes: Molecules which can act as catalysts in biological reactions.

F₁ Hybrid: The first generation progeny of the cross of two nonisogenicplants.

Genotype: The genetic constitution of a cell or organism.

Haploid: A cell or organism having one set of the two sets ofchromosomes in a diploid.

Linkage: A phenomenon wherein alleles on the same chromosome tend tosegregate together more often than expected by chance if theirtransmission was independent.

Marker: A readily detectable phenotype, preferably inherited incodominant fashion (both alleles at a locus in a diploid heterozygoteare readily detectable), with no environmental variance component, i.e.,heritability of 1.

Phenotype: The detectable characteristics of a cell or organism, whichcharacteristics are the manifestation of gene expression.

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer togenetic loci that control to some degree numerically representabletraits that are usually continuously distributed.

Resistance: As used herein, the terms “resistance” and “tolerance” areused interchangeably to describe plants that show no symptoms to aspecified biotic pest, pathogen, abiotic influence or environmentalcondition. These terms are also used to describe plants showing somesymptoms but that are still able to produce marketable product with anacceptable yield. Some plants that are referred to as resistant ortolerant are only so in the sense that they may still produce a crop,even though the plants are stunted and the yield is reduced.

Regeneration: The development of a plant from tissue culture.

Royal Horticultural Society (RHS) color chart value: The RHS color chartis a standardized reference which allows accurate identification of anycolor. A color's designation on the chart describes its hue, brightnessand saturation. A color is precisely named by the RHS color chart byidentifying the group name, sheet number and letter, e.g., Yellow-OrangeGroup 19A or Red Group 41B.

Self-pollination: The transfer of pollen from the anther to the stigmaof the same plant.

Single Locus Converted (Conversion) Plant: Plants which are developed bya plant breeding technique called backcrossing, wherein essentially allof the morphological and physiological characteristics of a spinachvariety are recovered in addition to the characteristics of the singlelocus transferred into the variety via the backcrossing technique and/orby genetic transformation.

Substantially Equivalent: A characteristic that, when compared, does notshow a statistically significant difference (e.g., p=0.05) from themean.

Tissue Culture: A composition comprising isolated cells of the same or adifferent type or a collection of such cells organized into parts of aplant.

Transgene: A genetic locus comprising a sequence which has beenintroduced into the genome of a spinach plant by transformation.

H. DEPOSIT INFORMATION

A deposit of spinach hybrid SV8545VC and inbred parent lines SMB66-1144Fand SMBS012-1188M, disclosed above and recited in the claims, has beenmade with the American Type Culture Collection (ATCC), 10801 UniversityBlvd., Manassas, Va. 20110-2209. The dates of deposit was Jul. 31, 2014.The accession number for those deposited seeds of spinach hybridSV8545VC and inbred parent lines SMB66-1144F and SMBS012-1188M are ATCCAccession No. PTA-121440, ATCC Accession No. PTA-121439, and ATCCAccession No. PTA-121438, respectively. Upon issuance of a patent, allrestrictions upon the deposits will be removed, and the deposits areintended to meet all of the requirements of 37 C.F.R. §1.801-1.809. Thedeposits will be maintained in the depository for a period of 30 years,or 5 years after the last request, or for the effective life of thepatent, whichever is longer, and will be replaced if necessary duringthat period.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity andunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the invention, as limited only bythe scope of the appended claims.

All references cited herein are hereby expressly incorporated herein byreference.

What is claimed is:
 1. A spinach plant comprising at least a first setof the chromosomes of spinach line SMB66-1144F or spinach lineSMBS012-1188M, a sample of seed of said lines having been depositedunder ATCC Accession Number PTA-121439 and ATCC Accession NumberPTA-121438, respectively.
 2. A seed comprising at least a first set ofthe chromosomes of spinach line SMB66-1144F or spinach lineSMBS012-1188M, a sample of seed of said lines having been depositedunder ATCC Accession Number PTA-121439 and ATCC Accession NumberPTA-121438, respectively.
 3. The plant of claim 1, which is an inbred.4. The plant of claim 1, which is a hybrid.
 5. The seed of claim 2,which is an inbred.
 6. The seed of claim 2, which is a hybrid.
 7. Theplant of claim 4, wherein the hybrid plant is spinach hybrid SV8545VC, asample of seed of said hybrid SV8545VC having been deposited under ATCCAccession Number PTA-121440.
 8. The seed of claim 6, wherein the seed isa seed of spinach hybrid SV8545VC, a sample of seed of said hybridSV8545VC having been deposited under ATCC Accession Number PTA-121440.9. A plant part of the plant of claim
 1. 10. The plant part of claim 9,wherein the plant part is a leaf, an ovule, pollen, a fruit, or a cell.11. A spinach plant having all the physiological and morphologicalcharacteristics of the spinach plant of claim
 7. 12. A tissue culture ofregenerable cells of the plant of claim
 1. 13. The tissue cultureaccording to claim 12, comprising cells or protoplasts from a plant partselected from the group consisting of embryos, meristems, cotyledons,pollen, leaves, anthers, roots, root tips, pistil, flower, seed andstalks.
 14. A spinach plant regenerated from the tissue culture of claim12, wherein said plant comprises all of the morphological andphysiological characteristics of the spinach plant comprising at least afirst set of the chromosomes of spinach line SMB66-1144F or spinach lineSMBS012-1188M, a sample of seed of said lines having been depositedunder ATCC Accession Number PTA-121439 and ATCC Accession NumberPTA-121438, respectively.
 15. A method of vegetatively propagating theplant of claim 1 comprising the steps of: (a) collecting tissue capableof being propagated from a plant according to claim 1; (b) cultivatingsaid tissue to obtain proliferated shoots; and (c) rooting saidproliferated shoots to obtain a rooted plantlet.
 16. The method of claim15, further comprising growing at least a first plant from said rootedplantlet.
 17. A method of introducing one desired trait into spinachline SMB66-1144F or SMBS012-1188M, said method comprising: (a) utilizingas a recurrent parent a plant of either spinach line SMB66-1144F orspinach line SMBS012-1188M, by crossing said plant of line SMB66-1144For SMBS012-1188M with a second donor spinach plant that comprises thedesired trait to produce F1 progeny, a sample of seed of said lineshaving been deposited under ATCC Accession Number PTA-121439, and ATCCAccession Number PTA-121438, respectively; (b) selecting an F1 progenythat comprises the desired trait; (c) backcrossing the selected F1progeny with a plant of the same spinach line used as the recurrentparent in step (a), to produce backcross progeny; (d) selecting abackcross progeny comprising the desired trait; and (e) repeating steps(c) and (d) three or more times to produce a selected fourth or higherbackcross progeny that comprises the desired trait, and otherwisecomprises all of the morphological and physiological characteristics ofthe recurrent parent spinach line used in step (a).
 18. A spinach plantproduced by the method of claim
 17. 19. A method of producing a plantcomprising an added trait, the method comprising introducing a transgeneconferring the trait into a plant of hybrid SV8545VC, line SMB66-1144For line SMBS012-1188M, a sample of seed of said hybrid and lines havingbeen deposited under ATCC Accession Number PTA-121440, ATCC AccessionNumber PTA-121439, and ATCC Accession Number PTA-121438, respectively.20. A plant produced by the method of claim
 19. 21. The plant of claim1, further comprising a transgene.
 22. The plant of claim 21, whereinthe transgene confers a trait selected from the group consisting of malesterility, herbicide tolerance, insect resistance, pest resistance,disease resistance, modified fatty acid metabolism, environmental stresstolerance, modified carbohydrate metabolism and modified proteinmetabolism.
 23. The plant of claim 1, comprising a single locusconversion.
 24. The plant of claim 23, wherein the single locusconversion confers a trait selected from the group consisting of malesterility, herbicide tolerance, insect resistance, pest resistance,disease resistance, modified fatty acid metabolism, environmental stresstolerance, modified carbohydrate metabolism and modified proteinmetabolism.
 25. A method for producing a seed of a plant derived from atleast one of hybrid SV8545VC, line SMB66-1144F or line SMBS012-1188Mcomprising the steps of: (a) crossing a spinach plant of hybridSV8545VC, line SMB66-1144F or line SMBS012-1188M with itself or a secondspinach plant; a sample of seed of said hybrid and lines having beendeposited under ATCC Accession Number PTA-121440, ATCC Accession NumberPTA-121439, and ATCC Accession Number PTA-121438, respectively; and (b)allowing a seed of the hybrid SV8545VC-derived, the lineSMB66-1144F-derived or the line SMBS012-1188M-derived spinach plant toform.
 26. The method of claim 25, further comprising the steps of: (c)selfing a plant grown from said hybrid SV8545VC-derived,SMB66-1144F-derived or SMBS012-1188M-derived spinach seed to yield anadditional hybrid SV8545VC-derived, line SMB66-1144F-derived or lineSMBS012-1188M-derived spinach seed; (d) growing said additional hybridSV8545VC-derived, line SMB66-1144F-derived or line SMBS012-1188M-derivedspinach seed of step (c) to yield an additional hybrid SV8545VC-derived,line SMB66-1144F-derived or line SMBS012-1188M-derived spinach plant;and (e) repeating the crossing and growing steps of (c) and (d) togenerate at least a first further hybrid SV8545VC-derived, lineSMB66-1144F-derived or line SMBS012-1188M-derived spinach plant.
 27. Themethod of claim 25, wherein the second spinach plant is of an inbredspinach line.
 28. The method of claim 25, comprising crossing lineSMB66-1144F with line SMBS012-1188M, a sample of seed of said lineshaving been deposited under ATCC Accession Number PTA-121439, and ATCCAccession Number PTA-121438, respectively.
 29. The method of claim 26,further comprising: (f) crossing the further hybrid SV8545VC-derived,SMB66-1144F-derived or SMBS012-1188M-derived spinach plant with a secondspinach plant to produce a seed of a hybrid progeny plant.
 30. A plantpart of the plant of claim
 7. 31. The plant part of claim 30, whereinthe plant part is a leaf, a flower, a fruit, an ovule, pollen, or acell.
 32. A method of producing a spinach seed comprising crossing theplant of claim 1 with itself or a second spinach plant and allowing seedto form.
 33. A method of producing a spinach fruit comprising: (a)obtaining a plant according to claim 1, wherein the plant has beencultivated to maturity; and (b) collecting a spinach fruit from theplant.